DE2552630A1 - Anti-eumycetes agent prepn. - from mixtures of japanese barberry and coexisting alkaloids plants - Google Patents

Abstract

Plants which contains alkaloids coexisting with alkaloids of berberine type or berberine series, or a plant except Japanese barberrys (Berberis thunbergii) which contains the alkaloids and the leaves of Japanese barberry are treated for removal of fat and tar, and extracted with a polar solvent. The extract is treated with a soln. of alkali metal dihydrogen phosphate and a soln. of phosphoric acid or organic acid, and then condensed. The condensate is then neutralised or slightly alkalised with a soln. of dialkali metal hydrogen phosphate or alkali metal phosphate (when dipotassium hydrogen phosphate is used in the first stage and disodium hydrogen phosphate used in the second stage, Ohbaku (Phellodendron amurense) is excluded). Water is removed, and the residue is extracted with a polar solvent. Removal of the solvent by distillation yields the anti-Eumycetes agent.

Description

Substance effective against fungi and process for their preparation

The invention relates to a substance for combating (controlling) real fungi, as well as a process for their preparation; it concerns in particular Means for combating (controlling) real fungi obtained from alkaloids of the berberine type containing plants are obtained, and a process for their production.

Among the berberine-type alkaloids are berberine, coptisine, worenin, palmatine and others, which are the common basic structure have to understand.

Only recently are effective and accurate methods for therapy of fungal diseases caused by pathogenic fungi such as Blastomyces, Candida, Cryptococcus, Coccidiodes, Epidermophyton, Histoplasma, Microsporum and Trichophyton be found. Antibiotics are currently used to treat these diseases of the polyene type such as amphotericin B, trichomycin and nystatin; these However, antibiotics have several disadvantages, e.g. B. the fact that they are toxic and their antifungal activity is uneven, and therefore they are not effective and targeted therapeutic effects.

The aim of the present invention is therefore to find substances for combating (Control) of real mushrooms (Fungi) indicate which have a high antifungal activity exhibit. The aim of the invention is also to provide a method for producing such indicate the real fungi-fighting (controlling) substances.

The invention relates to a method for producing a real Fungi-fighting (controlling) substance, which is characterized by that one contains a berberine-type alkaloid of fatty or resinous materials free plant tissues extracted with a polar solvent, the extract during or after extraction at a temperature of 35 to 750C with an aqueous, alkaline alkali dihydrogen phosphate solution brings the extract solution into contact bringing it into contact with the aqueous alkaline alkali metal dihydrogen phosphate solution constricts to form one on the extract saturated solution that thereby obtained saturated solution with an aqueous} alkaline dialkali hydrogen phosphate solution mixes to bring the pH of the mixture to 6.8 to 7.2, the polar solvent evaporated at a temperature of 35 to 75 ° C, the resulting residue under Reduced pressure dries the dried material with a polar solvent extracted and the extract solution evaporated to dryness.

The invention also relates to that produced by this method Product.

Examples of plants containing berberine-type alkaloids, are Philodendron amurense Ruprecht, P. amurense Rupr. var.

sachalinense, P amurense Rupr. var.japonicum (Maxim.) Ohwi and P. amurense Rupr. var. Levallei (Dode) Sprague, which belong to the family Rutaceae; Coptis japonica Makino, Coptis japonica Makino var.dissecta Nakoi, Coptis trifolia sabsbuty, Exanphorrhiza, and Xanphorrhiza simplicissima belonging to the Ranunculsceae family belong; Berberis Thunberzii, Berberis Sieboldí :, Berberis amurensis and Nandina domestica belonging to the Berberidaceae family; as well as Costinum fenestratum and Columbo Radix, which belong to the Menispermaceae family.

These plants contain berberine-type alkaloids in their tissues, such as B. Berberine, Palmatine, Coptesin, Worenin and the like., In the form of the hydroxide.

As the starting materials, those containing a berberine type alkaloid can be used Tissues of these plants are used, such as. B. the bark of which the cork layers been removed are, from Philodendron amurense Ruprecht, to the family belonging to the Rutaceae, the subterranean stems of Xanphorrhiza, to the family of Ranunculsceae belonging, the leaves, stems and roots of Berberis Thunberzii, Belonging to the Berberidaceae family, the stem of Costinum fenestratum, to Belonging to the Menispermaceae family and the leaves and stems of Coptis Rhizoma. In addition, dry powders of tissues of the above plants are commercially available, such as B. raw medicines under the names Philodendron powder, Coptis powder and the like, so that the use of these crude drugs is preferred.

The content of the starting materials containing a berberine-type alkaloid of resinous substances is preferably as little as possible. Accordingly the starting materials are preferably treated beforehand with a solvent, to the fatty, resinous or tarry substances contained therein remove. That is, the starting materials, i. H. the above plant tissues or raw medicinal products, are treated with a non-polar solvent, such as ether, Petroleum ether, mineral spirits, benzene, toluene, or xylene, are treated to help in this Remove solvent soluble components from it.

After drying, the extraction with the non-polar Solvent obtained residue with a suitable amount, for example the 2 to 3 times its volume, a polar solvent such as methanol, ethanol, Acetone, acetone alcohol and the like, further extracted. The extraction is carried out several times, preferably at a temperature of 35 to 75 C, carried out for so long until the last extract solution is practically colorless. The extract solution is filtered and with about 1/8 to 1/12 parts by volume of a 0.5 to 1.5 molar aqueous alkaline Alkalidihydrogenphosphatlösung added.

The mixture is heated to 35-75 ° C and the polar solvent is evaporated at this temperature under reduced pressure, one at the Extract saturated solution is obtained.

Extraction of the dried residue can be performed using a mixture of the polar solvent and the aqueous alkaline alkali metal dihydrogen phosphate solution in a mixing ratio of 5 to 25: 1. In this case can then from the extract solution obtained without further treatment with the aqueous alkaline alkali hydrogen phosphate solution immediately the polar solvent are evaporated to obtain the solution saturated with the extract.

The solution obtained in this way can after the complete Evaporation of the polar solvent under acidic conditions extracted with ether in order to remove further impurities from it, which dissolve in the ether.

The pH of the solution saturated with the extract is determined by using an alkaline solution of dialkali hydrogen phosphate, such as dikallum or disodium hydrogen phosphate, or by phosphoric acid and caustic alkali, brought to 6.8 to 7.2. The color of the mix becomes pale yellow-brown.

The mixture, which is a clear solution, is below evaporated under reduced pressure at a temperature of 800 c or less, wherein a concentrated solution is obtained which contains the desired substances, which can be dried under reduced pressure or by freeze drying.

The concentrated solution containing the desired substances can used as a means of combating (controlling) real fungi and after diluting administered to humans or animals with an appropriate amount of water. she can but also by repeatedly extracting the dried material with the polar ones Purified solvents and subsequent evaporation of the polar solvent yielding a more purified product. This purified product, which is a yellow-brown crystalline powder is described below referred to as substance "NT-72".

With the substance obtainable by the process according to the invention NT-72 it is possible with success the growth of various medicinal fungi, such as B. those belonging to the genera Aspergillus, Candida, Trichophyton and the like. belong to inhibit (to inhibit). By incorporating a phosphate buffer with The antifungal activity of the substance NT-72 can still have a pH value of 6.8 to 7.2 can be further improved and it can be stored in an extremely stable condition will. The substance according to the invention NT-72 is both in any amount miscible with hydrophilic as well as with oleophilic ointment base agents and they shows excellent Effect in the therapy of dermatophytosis in humans or animals, when applied to the skin in an amount of 0.2 to 0.5 g is applied to patients suffering from this disease.

It can help a patient suffering from internal fungi administered by the oral route at a dose of from about 0.01 to about 0.05 g / kg / day or it can be injected at a dose of from about 0.001 to about 0.005 g / kg / day will. The acute toxicity is so low that when administered orally to mice the LDSo of NT-72 is 7 g / kg; d. H. the toxicity is far less than that of berberine hydrochloride, the LD5Q of which is 60 to 70 mg / kg.

Although it has not yet been clarified which component attribute the antifungal activity of NT-72 to the berberine-containing plant is, it is believed that this product is a substance in of some of the berberine-type alkalis in a certain form with phosphoric acid are connected.

The substance NT-72 obtained from Philodendron melts between and 116 ° C and represents a combination of various components. When subjected to high-performance liquid high-speed chromatography, several maxima appear in the chromatogram. However, the active effectiveness in terms of growth inhibition in real fungi (mushrooms) can be seen in two maximum proportions, one of which is the most active.

The components of substance NT-72 are referred to below explained in more detail on the accompanying drawings.

Fig. 1 shows a chromatogram obtained when high-performance liquid chromatography using Nihon Bunko Co.'s FLC-350 device at one pressure of 30 kg / cm², at ambient temperature, at a flow rate of 1.6 ml / minute, a chart recording speed of 0.5 cm / minute using the column Jaskopack SD-C2 -500 and the detector UV-254 Uvidec-1,2-FP-4, RI became.

Fig. 1 shows 7 maxima. The maximum 1 goes to the remaining solvents which are water and alcohol. The maxima 2 to 7 correspond the components of NT-72. Each of the components is separated and by the disc method For their growth inhibiting activity on Aspergillus fumigatus, Candida albicans and Trichophyton mentagrophytes tested.

Table diameter of the circle of inhibition (mm) separated component 1 2 3 4 5 6 7 Aspergillus fumigatus (-) 7.0-7.2 (-) (-) 16-17 lane (-) lane Candida albicans (-) 11.5-12.4 (") 14-15 (-) (-) Trichophyton (-) 12.5-13.6 (-) (-) 28-29 Trace (-) mentagrophytes With the proviso that N-75 is an inorganic Contains salt of a berberine-type alkaloid as the active ingredient when passed through Reduction with sodium borohydride frees this inorganic acid and the substances obtained are extracted with various solvents, the most active component is extracted with butanol.

Now if berberine hydrochloride (Japanese Pharmacopeia Grade) as above is treated, the butanol extract obtained does not have the desired one Activity on. The results obtained are shown in Table II below specified.

Table II Diameter of the circle of inhibition (mm) NT-72 Aspergillus fumigatus Candida albicans methanol solution complete 10.0 incomplete 23> 0 23.0 aqueous solution 18.0 30.0 berberine hydrochloride methanol solution are (-) aqueous Solution (-) (-) In Fig. 2 are the infrared absorption spectra of the butanol extracts shown, which shows that the two substances are different from each other are.

3 and 4 each show NMR spectra of the butanol extracts of reduced NT-72 or reduced berberine hydrochloride, dissolved in heavy water. The common absorptions are indicated by the signal X and those of berberine missing absorptions are indicated by the signal.

5 shows the gas chromatograms of the butanol extracts of NT-72 and berberine hydrochloride. From this it can be seen that the maxima A, B, C and D are mutually exclusive are different and that the maxima 1, 2, 3, 4, 5 and 6 are common. The applied Gas chromatographic conditions were as follows: Column: Gas Chrom P Silicon GUM SE-30 0.75% 2.25 m Programming: 40 ° C to 240 ° C.

Speed: 400 / minute Chart recording speed: 5mm / minute Device: Janagimoto GCG-550 FP The invention is accomplished by the following Examples, experimental, reference and clinical examples explained in more detail, but without to be limited to it.

Example 1 freed 200 g of a powder from the bark of the cork layers / from Philodendrons were extracted several times with 500 ml of ether each time in order to remove the Remove any grease or resin materials it contains. The arrears were 2000 ml of ethanol and 25 ml of an aqueous 1M potassium dihydrogen phosphate (KH2P04) solution added and the resulting mixture was 2 to 3 hours at about 600 C treated, cooled and then filtered. The residue became the same 3 times Extraction process subjected as above.

Thereafter the filtrates were combined and the ethanol became by distillation under reduced pressure at below 60 0C from the combined The filtrate was removed to give a solution saturated with the extract. By The solution was added to an aqueous 1N disodium hydrogen phosphate (Na2HPO4) solution Made weakly alkaline (pH 7.2) and then freeze-dried. After freeze drying the residue was dissolved in 100 ml of ethanol and the resulting solution became filtered. Then the filtrate was reduced under reduced pressure at below 600C evaporated to dryness, 20 g of a yellow-brown crystalline powder contained.

The procedure described above was repeated, this time however, the ethanol in each case by the polar solvents ethanol and acetone alcohol was replaced to give the same result as above.

Example 2 200 g of a fine powder from which the cork layers have been removed Philodendron bark was extracted several times with 500 ml of petroleum ether to obtain to remove the fat and tar contained therein. About the residue 2000 ml of ethanol were added and the resulting mixture became 2 to 3 Heated to about 60 ° C for hours, cooled and then filtered.

The residue was subjected to the same extraction as above 3 times. Thereafter, the filtrates were combined and there were 25 ml of an aqueous 1M sodium dihydrogen phosphate solution was added. The mixture was reduced under Distilled to dryness at 60 ° C under pressure. The residue was dissolved in water and the resulting solution was 3 times each using about 1/3 their amount of ether shaken. The formed aqueous layer was separated, weak by adding an aqueous 1M disodium hydrogen phosphate (Na2 HP04) solution made alkaline (pH 7.2) and then freeze-dried. After freeze drying the residue was dissolved with 100 ml of ethanol and the resulting solution became filtered. The filtrate was then reduced to below 60 ° C. under reduced pressure evaporated to dryness and obtained 20 g of the same yellow-brown crystalline Powder as in Example 1.

The procedure described above was repeated, this time the ethanol is replaced by the polar solvents methanol and acetone to obtain the same result as above.

Bezuvsbeispiel (1) A powder freed from fat and tar, which on prepared in the same manner as in Example 1 from 200 g of a philodendron powder became 2000 ml of an ethanol solution containing 3% hydrochloric acid admitted. The resulting mixture was left to stand at room temperature overnight and then filtered and the filtrate was taken under reduced pressure at below Concentrated to 40 ° C. The residue was dissolved in water and the resulting solution was shaken 3 times using about 1/3 its amount of ether each time.

The aqueous layer thus formed was drawn off by addition made weakly alkaline (pH 7.2) and then a lOZOiaen aqueous ammonia solution freeze dried. After freeze-drying, the residue was dissolved in 100 ml of ethanol dissolved and the resulting solution was filtered. Then the filtrate was evaporated to dryness under reduced pressure at below 60 ° C. and obtained a yellowish-brown powder.

(2) Using 2000 ml of ethanol containing 3% tartaric acid, 200 g of a philodendron powder freed from fat and tar, which on the in the same manner as obtained in Example 1 as in the above section (1) indicated treated, whereby a yellowish-brown powder was obtained.

(3) Using 2000 ml of ethanol containing 3% acetic acid, 200 g of a tar and fat freed philodendron powder, based on the same Manner as in Example 1, as in section (1) above indicated treated, whereby a yellowish-brown powder was obtained.

0.02 g of each of those obtained in paragraphs (1) to (3) above yellowish-brown powder and the yellow-brown powder obtained in Examples 1 and 2 Crystalline powders were given orally to males twice a day for 2 weeks Mice with an average body weight of 20 to 22 g (average age 1.5 months). In the groups similar to those obtained in Examples 1 and 2 yellow-brown crystalline powder were administered, no dead mice were found observed (see the following Table III).

Table III Extraction Numbers of Mice Number of Living Numbers The procedure per group of mice after 2 weeks of dead wetness after 2 weeks of reference example (L) 10 2 8 (2) 10 7 3 (3) 10 6 4 Example 1 10 10 0 Example 2 10 10 0 Experiment example 1 Antifungal activity: 0.8 g of each of the yellow-brown obtained in Examples 1 and 2 crystalline powders were distilled in 1 ml of a phosphate buffer at pH 6.8, respectively Dissolved in water.

Using 0.01 ml of the resulting solution, the Growth-inhibiting effect of each powder on fungi (F (Fungi) by the disc method tested. There was an increase in the inhibitory effect in the case where the phosphate buffer was used, observed (see. The following Table IV).

Table IV Example 1 Example 2 Test Fungi Distilled pH 6.8 distilled pH 6.8 water water Trichophyton mentagrophytes 19 - 21 22-23 18 - 21 23-24 Candida albicans 19 - 21 23-24 18 - 20 22-24 Aspergillus fumigatus 19 - 21 23-24 19 - 21 23-24 [The numbers indicate the diameter of the inhibition circle (mm)] Experimental example 2 Aqueous solutions of nystatin, ainphotericin B and Byb 5097 and a phosphate buffer solution of the yellow-brown crystalline prepared in Example 1 Powders were each placed on an anti-fungi test disc (diameter 8 mm, weight 0.03 g), followed by 0.005 g of the active ingredient of each solution adsorb, and it became the inhibitory effect against Aspergillus fumigatus, Candida albicans, Trichophyton mentagrophytes and Cryptococcus neoformans were observed, being the results given in Table V below were obtained.

Table V in Example 1 amphoteric obtained ricin B nystatin Byb 5097 Powder Aspergillus 18-12 12.5 fumigatus (circular (lane 18) ge lane) Candida 25 11 21 17 albicans (lane 14) (lane 22) Trichophyton mentagro- 48.5 8.5 12.5 24 phytes Cryptococcus 11-11 neoformans (circular (lane 18) ge lane) [The numbers indicate the diameter of the inhibition circles (in mm)] Each fungi test liquid was mixed with a Sabouraud agar medlum containing 0.1% Twen 80 to form a multilayer fungi agar film which was then allowed to solidify. aside from that Amphotericin B, Nystatin and Byb 5097 in the phosphate buffer solution were insufficient soluble, however, it was believed to be sufficient in Abso + tion on the 8 mm disc had been incorporated in a sufficient amount.

Example 3 200 g of a Coptis powder were used 3 times with 500 ml of ether is extracted to remove the fatty or resinous materials it contains. The solid residue was dried and treated with 2000 ml of ethanol and the same The resulting mixture was allowed to stand at about 60 ° C. for 2 hours. To the mix became then 40 ml of an aqueous 1M potassium dihydrogen phosphate (KH2P04) solution was added the mixture was heated to about 600C, cooled and then filtered. The residue was subjected to the same procedure as above 3 times. After that, the filtrates were combined with one another and then at below 60 ° C a distillation under reduced Subjected to pressure. The residue was purified by adding an aqueous 1M disodium hydrogen phosphate (Na2HP04) solution made weakly alkaline (pH 7.2).

completely freed from ethanol by D2stRation and then freeze-rocked. After freeze-drying, the residue was dissolved in 100 ml of ethanol and the the resulting solution was filtered. Then the filtrate was reduced under reduced pressure Pressure evaporated to dryness at below 600C, giving 20 g of a yellow-brown crystalline powder received.

Example 4 200 g of a Coptis powder were obtained using Each 500 ml of ether extracted to the fat or resin materials contained therein to remove. To the residue were 2000 ml of ethanol and 56 ml of an aqueous 1M dipotassium hydrogen phosphate solution was added and the resulting mixture was Digested for 2 hours at about 600C, cooled and then filtered.

The residue was subjected to the same extraction as above 3 times. The filtrates were then combined with one another and the ethanol was desdEated removed from the combined filtrate under reduced pressure at below 60 ° C. The residue was dissolved in water and the resulting solution was used 3 times shaken using about 1/3 of their amount of ether. The formed aqueous Layer was separated by adding an aqueous 1M disodium hydrogen phosphate (Na2 HPO4) solution made weakly alkaline (pH 7.2) and then freeze-dried. After freeze-drying, the residue was dissolved in 100 ml of ethanol and the resulting resulting solution was filtered. Then the filtrate was reduced under reduced pressure The pressure was evaporated to dryness below 60 ° C. and 20 g of the same were obtained yellow-brown crystalline powder as in example 3.

The powder thus obtained was dissolved in 100 ml of a phosphate buffer (consisting of 40 ml M / 15 KH2PO4 and 60 ml M / 15 Na2HP04) dissolved to form a stable solution with a pH of 7.0. This solution was at for 6 months 300C, but with no change in terms of antifungal activity was observed.

Examples 5 to 10 Example 1 was repeated, but this time instead of the bark of philodendron in each case those given in the following table Vt Starting materials were used. The yields, the antifungal activities and the acute toxicity of the products is given in Table VI below.

The antifungal activity was determined according to that described in Experimental Example 1 Procedure determined. The acute toxicity was determined as follows: that in the examples The products obtained were in each case transferred into aqueous solutions, which then with Water were diluted in amounts of 65 to 70 mg / ml, 70 to 75 mg / ml, 75 to 80 mg / ml and 80 to 85 mg / mlX and the dilutions thereby obtained became the actual Subjected to toxicity test. The test was carried out in the manner that each test solution in the tails of 20 mice (male and female) per group (intravenous) was injected (85 mice were reserved) having a body weight from 21 to 24 g and an average body weight of 22.6 g, and the number of surviving mice was determined. The results obtained are given in Table VI below.

The numbers in Table VI represent the survival ratios of the mice represent; for example, the value 10/10 shows that 10 out of 10 mice survived, while the value 8.2 / 10 indicates that 8.2 out of 10 mice survived.

Table VI When- starting materials- from antifungal activity acute toxicity sp. rial (partial) prey Aspergil- Canadida Trichophy- 65 - 70 70 - 75 75 - 80 80 - 85 85 - 90 lus fumi- albicans ton menta- mg / ml mg / ml mg / ml mg / ml mg / ml No. (g) gatus grophytes 5 Costinum fenestratum (bark) 18.1 20-23 18-20 26-27 8.6 / 10 8/10 7.6 / 10 6/10 5/10 6 Berberis Thunberzii (leaves) 15.5 20-22 20-23 28-30 10/10 10/10 8.2 / 10 6.8 / 10 6/10 7 Barberis Thunberzii (stem) 20.2 23-24 21-23 28-32 10/10 8/10 7/10 6.2 / 10 6/10 8 Berberis Sieboldi (stems) 17.0 20-23 21-22 27-28 10/10 10/10 10/10 9/10 7/10 9 Philodendrom (bark) Barberis Thun- 17.5 18-19 18-19 24-25 9/10 7.6 / 10 6.4 / 10 5.2 / 10 5/10 berzii (leaves) each 100 10 Philodendron (bark) Col-umbo Radix 16.3 22-24 20-24 28-31 10/10 8/10 7.2 / 10 7/10 6/10 (fruits) 50: 150 Examples 11 and 12 Example 1 was repeated, this time using instead of the bark of Philodendron in each case the starting materials given in the following table and petroleum ether was used instead of ether. The yield, the antifungal activities and the acute toxicities of the products determined as in the examples above are given in Table VII below.

When- starting material- from antifungal activity acute toxicity sp. rial (Partial) prey Aspergil- Canadida Trichophy- 65 - 70 70 - 75 75 - 80 80 - 85 85 - 90 lus fumi- albicans ton menta- mg / ml mg / ml mg / ml mg / ml mg / ml No. (g) gatus grophytes 11 Costinum fenestratum (stems, roots) 18.2 22-24 21-23 28-30 8.7 / 10 8/10 7/10 5.6 / 10 5/10 12 Coptis powder 15.1 20-22 15-16 20-22 9/10 8.2 / 10 7710 5.6 / 10 5/10 Clinical Example A solution was prepared by adding 0.4 g of the in the example above 1 yellow-brown crystalline powdery substance obtained in 1 ml of a phosphate buffer have been resolved. This solution was used as the starting liquid (final pH 7.2).

The starting liquid was diluted 50 times and 30 ml the dilution (0.24 g of the yellow-brown powdery substance) were 3 times am Days 3 to 3 weeks by oral route are those given in Table VIII below Given to patients with fungi pneumonia or urinary tract infection, being the increase or decrease in the number of fungi and bacteria in the sputum and Urine and the fluctuation in the concentration of alkaloids in the blood were observed. The results obtained are shown in Table VIII below.

As can be seen from Table VIII, the amounts of fungi and Bacteria in the sputum and urine decreased or became o with increasing concentration of the product according to the invention in the blood and the yellow-brown according to the invention powdered substance proved to be clinically effective not only against fungi, but also against bacterial infectious diseases caused by Pseudomonas aeruginosa, Escherichia coli and Micrococcus pyogenes. aside from that the therapeutic effects were observed, the lowering of fever and accompanied the decrease or disappearance of the chest shadows on the X-ray. the Administration of this drug did not result in any impairment of functions the liver and kidneys and no other side effects.

Table VIII Name the initial value first week second week third fourth Illness week week Parkinson's bacteria - in sputum (number of cells) (full test Candida 7000 # 80 # 55 # 3000 Stiffness) in the urine # 59 years old Escherichia 300 # 400 # 220 # (-) Concentration (2 hours after administration in the blood) 0.12 (# / ml) 8 18 1.5 Administration Duration # 14 days # Fungi-Men in Bacteria- in Sputum gitis test Pseudomonas 2000 (Asperigillus Candida 800 Infection) in the urine # 41 years old Candida 600 # 40000 # 500 Escherichia 3000 # 6000 # 4500 Concentration (2 hours after the 14 in the blood) 0.4 (# / ml) Administration duration # 7 days # Table VIII continued Name the initial value first second third fourth Disease week week week week Backbone damage bacteria test in sputum # 27 years as Pseudomonas 50000 # 80000 in the urine 140 # 200 # 400 # 10 Staphylococcus Concentration (2 hours after administration in the blood chung) + (# / ml) 8.8 5 Administration duration # 7 days # Dislocation bacteria test in sputum # 39 years old Pseudomonas 140000 # 140 # 20 # (-) in the urine Pseudomonas 600 # 600 # 1000 # 600 Staphylococcus 600 Escherichia 400 Concentration (2 hours after administration in the blood) + (# / ml) 2.4 2.2 25 Administration duration # 21 days # Rupture of the cerebral bacteria test in the sputum Candida aneurysm 160000 # 4000 # 8000 # 7000 # 30 # 67 years old in the urine Pseudomonas 60000 # 5000 # 1000 # 60000 # 7000 Candida 200 Table VIII continued Name the initial value first second third fourth Disease week week week week Concentration (3 hours after administration in the blood chung) 0.5 (# / ml) 2.2 18 0.8 40 Administration Duration # 14 days # # 7 days # Cerebral bacteria test in the spectrum Thrombosis Candida 40000 # 40 # 30000 # 3000 # (-) # 59 years Escherichia 20,000 350 old in the urine Escherichia 8000 # (-) Candida 460 # (-) Concentration (3 hours after administration in the blood chung) + (# / ml) 8 1.8 12 16 Administration Duration # 28 days # Cerebral backterial test in sputum Thrombosis Candida 100000 # 20 # 400 # 200 # 59 years of Micrococcus 8000 # 8000 # 400 old in urine Candida 400 # 20 # (-) # (-) Staphylococcus 140 # (-) # (-) Concentration (3 hours after administration in the blood chung) + (# / ml) 24 16 16 Administration duration # 21 days # Footnotes: (1.) Candida: Candida albicans Pseudomonas: Pseldomonas aeruginosa Escherichia: Escherichia coli Saphylococcus: Staphylococcus aureus (2.) The bacteria test was carried out every 7 days.

The measurement of the concentration in the blood was 2 bis for the first time 3 hours after the administration of the drug and then every 7 days at the same time carried out with the bacterial test.

(3.) The alkaloid concentration in the blood was determined by 2 methods: a method using a Wagner reagent and a method for determination the antifungal value of the serum against Aspergillus fumigatus (100,000 / ml).

The invention has been preferred with reference to z-ar above Embodiments explained in more detail, but it is a matter of course for the Facnmann, that it is by no means restricted to this, but that it is in many respects Ennen are changed and modified without thereby departing from the scope of the present Invention is abandoned.

Claims L e r s e i t e

Claims (17)

Claims tL? Process for the preparation of a substance for Control (control) of data fungi, characterized in that an alkaloid Berberine-type plant tissues free of adipose or resinous materials extracted with a polar solvent, the extract during or after the extraction at a temperature of 35 to 750 ß with an aqueous alkaline alkali dihydro gene phosphate solution brings into contact, the extract solution after contacting with the aqueous alkaline alkali metal dihydrogen phosphate solution constricts with formation a solution saturated with the extract, the saturated solution obtained with it an aqueous alkaline dialkali hydrogen phosphate solution is mixed to the pw value bring the mixture to 6.8 to 7.2, the polar solvent at a temperature evaporated from 35 to 75 ° C, the resulting residue under reduced pressure dried, the dried material extracted with a polar solvent and the polar solvent evaporates to dryness. -2. Method according to claim 1, characterized in that times as a plant tissue containing an alkaloid of the berberine type a tissue from philodendron, Coptis, Costinum, Co1umbo and Berberis are used. 3. The method according to claim 1 and / or 2, characterized in that that times as a plant tissue containing a berberine type alkaloid, raw medicines containing or consisting of philodendron powder or coptis powder. 4. The method according to at least one of claims 1 to 3, characterized characterized in that one plant tissue containing an alkaloid of the berberine type used from which the fat or resin materials have been removed by extraction the fatty or resinous materials made from those containing a berberine-type alkaloid Plant tissues using a non-polar solvent from the ether group, Petroleum ether, petroleum ether, benzene, toluene and xylene. 5. The method according to at least one of claims 1 to 4, characterized characterized in that one is used as a polar solvent for extracting the an alkaloid Berberine-type plant tissue that is free from fatty or resinous materials > methyl alcohol, ethyl alcohol, acetone or acetone alcohol is used. 6. The method according to at least one of claims 1 to 5, characterized characterized by extracting the containing an alkaloid of the barberine type Plant tissue at a temperature of 35 to 75 ° C. 7. The method according to claim 6, characterized in that the Repeated extraction several times. 8. The method according to at least one of claims 1 to 7, characterized characterized in that one uses a 0.5 to 1.5 molar aqueous alkaline alkali metal dihydrogen phosphate solution used. 9. The method according to at least one of claims 1 to 8, characterized characterized in that the aqueous alkaline Alkali dihydrogen phosphate solution, which is brought into contact with the extract, used in such a close manner that about 1/8 to 1/12 the volume of the polar used to extract the tissue Solvent corresponds. 10. The method according to at least one of claims 1 to 9, characterized characterized in that the concentration of the extract solution by evaporating the polar Carries out solvent under reduced pressure at a temperature of 35 to 75 ° C. 11. The method according to at least one of claims 1 to 10, characterized characterized in that the alkaline alkali metal dihydrogen phosphate is potassium dihydrogen phosphate or sodium di-hydrogen phosphate is used. 12. The method according to at least one of claims 1 to 11, characterized characterized in that the alkaline dialkali metal hydrogen phosphate is dipotassium hydrogen phosphate or disodium hydrogen phosphate is used. 13. The method according to at least one of claims 1 to 12, characterized characterized in that the residue is dried by freeze drying. 14. The method according to at least one of claims 1 to 13, characterized characterized in that the polar solvent with which the residue obtained is used is extracted, ethyl alcohol is used. 15. Substance for the control (control) of real fungi, thereby characterized in that it is made according to the method according to at least one of the claims 1 to 14 has been produced. 16. Substance according to claim 15, characterized in that it contains or consists of a mixture of phosphoric acid Bound berberine-type alkaloids obtained after performing high-performance liquid chromatography from the Y, (constituent substance NT-72 exists. 17. Means for combating (controlling) real fungi, characterized in that that it contains or consists of the substance according to claim 15 and / or 16 and phosphates, which result in a buffer solution with pH 6.8 to 7.2.